Nodule RNAseq from M. truncatula NPD knockout lines

A nodule-specific subset of PLAT domain proteins (NPDs) expanded to 5 members in the Medicago lineage. In order to characterize NPD gene function, a set of cumulative NPD knockout lines was generated. A two-gene knockout line npd2/4, three-gene knockout lines npd1/2/4 and npd2/4/5, and wild type M. truncatula R108 were inoculated with Ensifer meliloti strain 1021. Plants were grown in a growth chamber at 22C and with a 16 h photoperiod in conetainers arranged in a completely randomized design. Trays were watered with sterile deionized water twice a week and with Fahräeus medium lacking nitrogen once a week. Thirty days post inoculation, roots and nodules were harvested and flash-frozen in liquid nitrogen. Total RNA was extracted with a Qiagen RNeasy Plant Mini Kit and sequenced by the University of Minnesota Genomics Center.